Team:Paris Saclay/Notebook/July/16

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Notebook : July 16

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining Bba_K1155003, Bba_K1155007

1 - Extraction of Bba_B0015, Bba_B0017, Bba_I732019 from DH5α

Zhou

Protocol : High copy plamid extraction

1 - Digestion of Bba_B0015, Bba_B0017, Bba_I732019 by EcoRI/PstI

Anaïs

  • DNA : 5µL
  • Buffer FD : 2µL
  • EcoRI FD : 1µL
  • PstI FD : 1µL
  • H2O : 21µL

We let our digestion 1h30 at 37°C.

Objective : obtaining biobricks in PSB3K3

1 - Digestion of PSB3K3 by EcoRI/PstI

Anaïs

Used quantities :

  • DNA : 5µL
  • Buffer FD : 2µL
  • EcoRI FD : 1µL
  • PstI FD : 1µL
  • H2O : 11µL

We let our digestion 1h30 at 37°C.

2 - Electrophoresis of the digestion of PSB3K3 by EcoRI/PstI

Sheng

[[]]
  • Well 1 : 5µL of PSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye
  • Well 2 : 5µL of PSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye
  • Well 3 : 6µL of DNA ladder
  • Well 4 : 5µL of PSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye
  • Well 5 : 5µL of PSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye
  • Gel : 1%

Expected sizes :

  • PSB3K3 : 2750bp

We obtain fragments at the right size.

3 - Gel purification of electrophoresis of the digestion of PSB3K3 by EcoRI/PstI

Abdou

Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]

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